Degradation Pathways Of Atrazine Research Articles

Experimental in situ chemical peroxidation of atrazine in contaminated soil

Lab-scale experiments of in situ chemical oxidation (ISCO), were performed on soil contaminated with 100 mg kg −1 of atrazine (CIET). The oxidant used was hydrogen peroxide catalysed by naturally occurring minerals or by soluble Fe(II) sulphate, added in aqueous solution. The oxidation conditions were: CIET:H 2O 2 = 1:1100, 2 PV or 3 PV reaction volume, Fe(II):H 2O 2 = 0, 1:22, 1:11. Stabilized (with KH 2PO 4 at a concentration of 16 g l −1) or non-stabilized hydrogen peroxide was used. The pH of the reagents was adjusted to pH = 1 with sulphuric acid, or was not altered. Results showed that the addition of soluble Fe(II) increased the temperature of the soil slurry and the use of stabilized hydrogen peroxide resulted in a lower heat generation. The treatment reduced the COD of the soil of about 40%, pH was lowered and natural organic matter became less hydrophobic. The highest atrazine conversion (89%) was obtained in the conditions: 3 PV, Fe(II):H 2O 2 = 1:11 with stabilized hydrogen peroxide added in two steps. The stabilizer only increased H 2O 2 life-time significantly when soluble Fe(II) was added. Results indicate as preferential degradation pathway of atrazine in soil dechlorination instead of dealkylation.

Simultaneous degradation of atrazine and phenol by Pseudomonas sp. strain ADP: effects of toxicity and adaptation.

The strain Pseudomonas sp. strain ADP is able to degrade atrazine as a sole nitrogen source and therefore needs a single source for both carbon and energy for growth. In addition to the typical C source for Pseudomonas, Na(2)-succinate, the strain can also grow with phenol as a carbon source. Phenol is oxidized to catechol by a multicomponent phenol hydroxylase. Catechol is degraded via the ortho pathway using catechol 1,2-dioxygenase. It was possible to stimulate the strain in order to degrade very high concentrations of phenol (1,000 mg/liter) and atrazine (150 mg/liter) simultaneously. With cyanuric acid, the major intermediate of atrazine degradation, as an N source, both the growth rate and the phenol degradation rate were similar to those measured with ammonia as an N source. With atrazine as an N source, the growth rate and the phenol degradation rate were reduced to approximately 35% of those obtained for cyanuric acid. This presents clear evidence that although the first three enzymes of the atrazine degradation pathway are constitutively present, either these enzymes or the uptake of atrazine is the bottleneck that diminishes the growth rate of Pseudomonas sp. strain ADP with atrazine as an N source. Whereas atrazine and cyanuric acid showed no significant toxic effect on the cells, phenol reduces growth and activates or induces typical membrane-adaptive responses known for the genus Pseudomonas. Therefore Pseudomonas sp. strain ADP is an ideal bacterium for the investigation of the regulatory interactions among several catabolic genes and stress response mechanisms during the simultaneous degradation of toxic phenolic compounds and a xenobiotic N source such as atrazine.

Nitrogen control of atrazine utilization in Pseudomonas sp. strain ADP.

Pseudomonas sp. strain ADP uses the herbicide atrazine as the sole nitrogen source. We have devised a simple atrazine degradation assay to determine the effect of other nitrogen sources on the atrazine degradation pathway. The atrazine degradation rate was greatly decreased in cells grown on nitrogen sources that support rapid growth of Pseudomonas sp. strain ADP compared to cells cultivated on growth-limiting nitrogen sources. The presence of atrazine in addition to the nitrogen sources did not stimulate degradation. High degradation rates obtained in the presence of ammonium plus the glutamine synthetase inhibitor MSX and also with an Nas(-) mutant derivative grown on nitrate suggest that nitrogen regulation operates by sensing intracellular levels of some key nitrogen-containing metabolite. Nitrate amendment in soil microcosms resulted in decreased atrazine mineralization by the wild-type strain but not by the Nas(-) mutant. This suggests that, although nitrogen repression of the atrazine catabolic pathway may have a strong impact on atrazine biodegradation in nitrogen-fertilized soils, the use of selected mutant variants may contribute to overcoming this limitation.

Purification, substrate range, and metal center of AtzC: the N-isopropylammelide aminohydrolase involved in bacterial atrazine metabolism.

N-Isopropylammelide isopropylaminohydrolase, AtzC, the third enzyme in the atrazine degradation pathway in Pseudomonas sp. strain ADP, catalyzes the stoichiometric hydrolysis of N-isopropylammelide to cyanuric acid and isopropylamine. The atzC gene was cloned downstream of the tac promoter and expressed in Escherichia coli, where the expressed enzyme comprised 36% of the soluble protein. AtzC was purified to homogeneity by ammonium sulfate precipitation and phenyl column chromatography. It has a subunit size of 44,938 kDa and a holoenzyme molecular weight of 174,000. The K(m) and k(cat) values for AtzC with N-isopropylammelide were 406 micro M and 13.3 s(-1), respectively. AtzC hydrolyzed other N-substituted amino dihydroxy-s-triazines, and those with linear N-alkyl groups had higher k(cat) values than those with branched alkyl groups. Native AtzC contained 0.50 eq of Zn per subunit. The activity of metal-depleted AtzC was restored with Zn(II), Fe(II), Mn(II), Co(II), and Ni(II) salts. Cobalt-substituted AtzC had a visible absorbance band at 540 nm (Delta epsilon = 84 M(-1) cm(-1)) and exhibited an axial electron paramagnetic resonance (EPR) signal with the following effective values: g((x)) = 5.18, g((y)) = 3.93, and g((z)) = 2.24. Incubating cobalt-AtzC with the competitive inhibitor 5-azacytosine altered the effective EPR signal values to g((x)) = 5.11, g((y)) = 4.02, and g((z)) = 2.25 and increased the microwave power at half saturation at 10 K from 31 to 103 mW. Under the growth conditions examined, our data suggest that AtzC has a catalytically essential, five-coordinate Zn(II) metal center in the active site and specifically catalyzes the hydrolysis of intermediates generated during the metabolism of s-triazine herbicides.

Characterization of an atrazine-degrading Pseudaminobacter sp. isolated from Canadian and French agricultural soils.

Atrazine, a herbicide widely used in corn production, is a frequently detected groundwater contaminant. Fourteen bacterial strains able to use this herbicide as a sole source of nitrogen were isolated from soils obtained from two farms in Canada and two farms in France. These strains were indistinguishable from each other based on repetitive extragenic palindromic PCR genomic fingerprinting performed with primers ERIC1R, ERIC2, and BOXA1R. Based on 16S rRNA sequence analysis of one representative isolate, strain C147, the isolates belong to the genus Pseudaminobacter in the family Rhizobiaceae. Strain C147 did not form nodules on the legumes alfalfa (Medicago sativa L.), birdsfoot trefoil (Lotus corniculatus L.), red clover (Trifolium pratense L.), chickpea (Cicer arietinum L.), and soybean (Glycine max L.). A number of chloro-substituted s-triazine herbicides were degraded, but methylthio-substituted s-triazine herbicides were not degraded. Based on metabolite identification data, the fact that oxygen was not required, and hybridization of genomic DNA to the atzABC genes, atrazine degradation occurred via a series of hydrolytic reactions initiated by dechlorination and followed by dealkylation. Most strains could mineralize [ring-U-(14)C]atrazine, and those that could not mineralize atrazine lacked atzB or atzBC. The atzABC genes, which were plasmid borne in every atrazine-degrading isolate examined, were unstable and were not always clustered together on the same plasmid. Loss of atzB was accompanied by loss of a copy of IS1071. Our results indicate that an atrazine-degrading Pseudaminobacter sp. with remarkably little diversity is widely distributed in agricultural soils and that genes of the atrazine degradation pathway carried by independent isolates of this organism are not clustered, can be independently lost, and may be associated with a catabolic transposon. We propose that the widespread distribution of the atrazine-degrading Pseudaminobacter sp. in agricultural soils exposed to atrazine is due to the characteristic ability of this organism to utilize alkylamines, and therefore atrazine, as sole sources of carbon when the atzABC genes are acquired.

Atrazine Degradation in Irradiated Iron/Oxalate Systems: Effects of pH and Oxalate

The purpose of this study was to examine the various factors that control the kinetics of atrazine degradation in irradiated Fe(III)/oxalate systems, in the following denoted as photo-Fenton systems. In these systems, attack by hydroxyl radicals (HO•) was the only pathway of atrazine degradation. Transformation of pollutants by HO• that are produced in photo-Fenton systems is of interest in atmospheric waters, in iron-rich surface waters, and possibly on soil surfaces. Studies were conducted in systems containing 6 μM iron and 0, 18, and 180 μM oxalate at 3 ≤ pH ≤ 8, irradiated with simulated sunlight. Both oxalate concentration and pH greatly affected the rate of atrazine transformation. In the presence of initial 18 μM oxalate, the rate increased in the order of pH 7.5 7. In the absenc...

Modifications to atrazine degradation pathways in a loamy soil after addition of organic amendments

The effects of two organic amendments, a municipal solid waste compost and a composted straw, on [U- ring- 14C]atrazine degradation pathways in a loamy soil (Grignon, Yvelines, France) were studied during laboratory incubations under controlled conditions. Three month conditionings were conducted under sterile or non-sterile conditions, with soil plus atrazine, organic amendment plus atrazine, or soil plus organic amendment. Then either an organic amendment or soil or atrazine was added, respectively, to these three treatments and incubated for an additional 3 months under non-sterile conditions. Both organic amendments modified the behaviour of atrazine in soil but via different processes. The addition of municipal compost increased atrazine sorption and decreased its availability for degradation by soil microorganisms. The effect of the composted straw was mainly related to its high enzymatic activity, which appeared to be responsible for the production of large amounts of hydroxyatrazine. This hydroxylation favoured the opening of the triazine ring and its subsequent mineralization in the soil. At the end of the incubations, less atrazine was mineralized in the presence of the two types of organic amendment, which both increased the formation of non-extractable residues of atrazine. The addition of municipal compost preserved larger amounts of extractable atrazine, while the addition of composted straw enhanced hydroxyatrazine production. In all cases, the greatest effects were found when atrazine was directly in contact with the organic amendment during conditioning.

The atzB gene of Pseudomonas sp. strain ADP encodes the second enzyme of a novel atrazine degradation pathway.

We previously reported the isolation of a 21.5-kb genomic DNA fragment from Pseudomonas sp. strain ADP, which contains the atzA gene, encoding the first metabolic step for the degradation of the herbicide atrazine (M. de Souza, L. P. Wackett, K. L. Boundy-Mills, R. T. Mandelbaum, and M. J. Sadowsky, Appl. Environ. Microbiol. 61:3373-3378, 1995). In this study, we show that this fragment also contained the second gene of the atrazine metabolic pathway, atzB. AtzB catalyzed the transformation of hydroxyatrazine to N-isopropylammelide. The product was identified by use of high-performance liquid chromatography, mass spectrometery, and nuclear magnetic resonance spectroscopy. Tn5 mutagenesis of pMD1 was used to determine that atzB was located 8 kb downstream of atzA. Hydroxyatrazine degradation activity was localized to a 4.0-kb ClaI fragment, which was subcloned into the vector pACYC184 to produce plasmid pATZB-2. The DNA sequence of this region was determined and found to contain two large overlapping divergent open reading frames, ORF1 and ORF2. ORF1 was identified as the coding region of atzB by demonstrating that (i) only ORF1 was transcribed in Pseudomonas sp. strain ADP, (ii) a Tn5 insertion in ORF2 did not disrupt function, and (iii) codon usage was consistent with ORF1 being translated. AtzB had 25% amino acid identity with TrzA, a protein that catalyzes a hydrolytic deamination of the s-triazine substrate melamine. The atzA and atzB genes catalyze the first two steps of the metabolic pathway in a bacterium that rapidly metabolizes atrazine to carbon dioxide, ammonia, and chloride.

Degradation pathways of atrazine under solar light and in the presence of TiO 2 colloidal particles

The transformation of atrazine under simulated solar light in the presence of particulates of titanium dioxide has been shown to occur through different routes: (i) hydrolysis of the 2-chloro substituent; (ii) alkyl chain oxidation; (iii) dealkylation of the side chain and final displacement of the amino with hydroxy groups. A series of degradation intermediates have been identified by means of chromatographic and mass spectrometric techniques. Cyanuric acid (e.g. 2,4,6-trihydroxy-1,3,5-triazine) is found not to be susceptible of further fast decomposition and can be considered as the final product of photodegradation. Comparison with other photolytic processes (e.g. direct, sensitized, in the adsorbed phase on alumina, in the presence of humic materials) is presented.

Identification of a Suitable Solid Carrier as a Matrix for the Permeable Reactive Barrier of Laboratory Scale Model Aquifer for Atrazine Degradation

Three different types of physical substrata such as sand, granualar activated carbon (GAC) and plastic macaroni beads were selected to identify a suitable solid carrier as a course matrix for the permeable reactive barrier (PRB) in a sand filled two-dimensional laboratory-scale model aquifer. An adhesion experiment was performed and tested with Pseudomonas sp ADP (PASP) under agitated as well as static conditions. In static conditions, adhesion to GAC was the highest (80%) followed by that to beads (60%). No adhesion to sand was observed under static or agitated conditions. Since atrazine is adsorbed by GAC, its bioavailability for degradation by PADP may be affected. To test this, simultaneous loss of atrazine and release of chloride (dechlorination, the first step in the pathway of atrazine degradation by PADP) were determined in the presence and absence of GAC. Although cells remained viable, no significant chloride release was detected. Thus, plastic macaroni beads were identified and selected as the most suitable support for PADP in the biological permeable reactive barrier (BPRB) of the model aquifer.Key words: Suitable Solid; Laboratory Scale; PADPTribhuvan University JournalVol. XXV, No. 1, 2005Page:119-128Uploaded date: 26, September, 2010

Atrazine Hydrolysis in Soil

AbstractAn important pathway of atrazine degradation in perfusion systems of three soils was chemical hydrolysis to hydroxyatrazine. Ultraviolet spectrophotometric analyses of the perfusates showed the presence and accumulation of hydroxyatrazine. Atrazine degradation followed first‐order kinetics in soil‐free, sterilized soil and perfusion systems. An increased rate of atrazine hydrolysis in an acid soil was consistent with the effect of pH on hydrolysis. No microbial degradation of atrazine was detected following inoculation of a soil‐free atrazine medium with perfusates. An increased rate of hydrolysis in the presence of sterilized soil was postulated to result from soil adsorption of atrazine. Soil pH and organic matter content largely controlled the rate of atrazine hydrolysis; for soils of similar pH, atrazine degradation rates increased with increased atrazine adsorption.